ENZYMES 1. what is an enzyme: -specific proteins that catalyze biochemical reactions - is not consumed by the reaction 2.

what are the six classes of enzyme and the reaction type each catalyzes? - OXIDOREDUCTASEox-reduction rxns - TRANSFERASEcatalyze the transfer of a group other then H from one substrate to another - HYDROLASEcatalyze hydrolysis of ether , ester, C-C, and P-N bonds - LYASE...catalyze removal of groups without hydrolysis and result in double bonds - ISOMERASEcatalyze interconversion of geometric, optical, or positional isomers - LIGASEcatalyze the joining of two substrates coupled with breaking of a pyrophoshate bond in ATP 3. which 5 enzymes are important in diagnosis of liver disease? - ALT AST LD GT

ALP

4. what are the substrates and products in reactions catalyzed by ALT and what are the major tissue sources? - Substrates are amino acids L-alanine & L-glutamate - The products are alpha keto acids alpha ketoglutarate, and pyruvate - Major tissue sources are mainly liver and some kidney 5. what are the substrates and products in reactions catalyzed by AST and what are the major tissue sources? - Substrates are amino acids L-aspartate and L-glutamate - Products are alpha keto acids aa-ketoglutarate and oxaloaacetate - Major tissue sources are the heart and liver, some skeletal 6. what are the substrates and products in reactions catalyzed by LD and what are the tissue sources? - Substrates and products are lactate and pyruvate - Sources are heart, liver, skeletal muscle, kidney and rbc 7. what is the substrate and product in reactions catalyzed by ALP and what are the major tiss sources? - Substrate is p-nitrophenylphosphate (p-NPP) - Product is p-nitrophenol (p-NP) - Sources are liver and bone - Some in spleen, kidney, intest, placenta, renal tiss

8. what are the substrates and products in reactions catalyzed by CK and what re the major tiss sources - substrates are ATP and creatine phoshate - products are ADP and creatine - sources are skeletal muscle (highest activity), heart, brain, kidney & diaphragm 9. what are the substrates and products in reactions catalyzed by ACHE and what are the major tiss sources? - Substrates are acetylcholine - Products are choline and aacetic acid - Sources include nerve endings (major), rbc, lungs, spleen, gray matter of brain 10. what are the substrates and products in reactions catalyzed by lipase and what are th major tiss sources - substrates are ester linkages of triglycerides - products are glycerol and fatty acids - sources include pancrease (primary souce), and tongue 11. what are the major tissue sources of ACP? - Prostate gland (richest source), liver, spleen, bone marrow, platelets 12. what are the major tissue sources of amylase? - Pancrease (primary) - Saliva (secondary) 13. what are the major tissue sources of Sche - serum (major) - pancrease - liver (major) - heart - white matter of the brain

14. what is the clinical significance of elevated ALT? - Increased ALT activity is primarly liver specific and rarely observed in other conditions - Indicates that there is a problem with the liver 15. what is the clinical significance of elevated AST? - Elevated AST results are diagnostic for specific tissue damage only when compared with other measured enzyme activities (ieALT, CK) - Cardiac and liver disease - Progressive muscular dystrophy - Dermatomyositis - Muscle trauma, gangrene, pancreatitis, hemolytic disease - Cerebro vascular accidents, brain or spinal malignancies can cause increased AST in CSF

16. an ALT\/AST ratio of >1 is indicative of what? - Necrosis of the liver tissue 17. an ALT/AST ratio of <1 is indicative of what? - Cirrhosis and obstructive liver disease 18. what is the historical name of ALT? - SGPT 19. what is the historical name of AST? - SGOT 20. in the analysis of ALT what product is formed, what is measured, and at what wavelength? - The product pyruvate is reduced to lactate with the simultaneous oxidation of NADH - The oxidation of NADH to NAD - 340nm 21. in the analysis of AST, what product is formed, what is measured, and at what wavelength? - Oxaloacetate formed is reduced to malate by malate dehydrogenase (MDH) with the simultaneous oxidation of NADH - The oxidation of NADH to NAD - 340 nm 22. what functions does GT serve in the body? - Transport of amino aacids thru cell membrane 23. what enxyme is the most sensitive indicator of hepatobiliary disease? - GT 24. alcohol induced cirrhosis of the liver would be most indicated by what enzyme? - GT 25. what is the specimen of choice for enzymatic analysis? - Serum free of hemolysis - EDTA plasma can be used 26. in the kintic method for GT analysis, what is the color produced and what wavelength is used for analysis? - Yellow color - 405 nm 27. what function does LD serve in tissue when oxygen is limited?

LD facilitates the reoxidation of NADH coupled with the reduction of pyrubate to lactatenow glycolysis can continue

28. what is an aisoenzyme? - Different forms of an enzyme that are differentiated by variations in physical properties. 29. what are the 5 LD isoenzymes and the source of each? - LD1 (HHHH)heart - LD2 (HHHM)kidney brain rbc lung heart - LD3 (HHMM)brain, rbc, lung and spleen - LD4 (HMMM)brain, spleen - LD5 (MMMM)liver 30. name the best method for isoenzyme separation? - Electrophoresis 31. total LD enzyme increase, an isomorphic pattern, ( increase in all isoenzymes) is indicative of what? - Observed in multisystem diseases or injuries i. Hypoxia, extreme hyperthermia, congestive heart failure 32. what is the clinical significance of elevated LD1, LD2, LD3, LD4& LD5? - LD1 & LD2---hemolysis, anemias, renal disease or tumors, germ cell tumors, MI - LD1/LD2 flip---MI, pernicious anemia, renal infarction - LD2, LD3, LD4massive platelet destruction, transfusions, pulmonary embolism, lymphatic system involvement(infectious mono, lymphomas, lymphocytic leukemia - LD5liver disease (cirrhosis, hepatitis, passive congestion, liver carcinomas) skeletal muscle (extreme exercise or crushing injuries, progressive muscular dystrophy) 33. CSF LD activity is used for differential diagnosis of what disease? - Used for diff diagnosis of bacterial or viral meningitis - High LD in 90% bacterial 34. an elevated ALP level is indicative of what? - Bone disease (highest activity) - Hepatobiliary disease (2.5 times the upper limit) - Hyperparathyroidism (sl to mod increase) - Hypophospatasis----decreased serum activity 35. what 3 enzymes are good indicators of cardiac disease? - CK (CK-MB), LD, AST 36. CKs major function is?

Catalyze reversible phosphorylation between creatine and creatine phosphate yielding ATP for muscle metabolism

37. name the CK isoenzymes and their subunit structures - CK-1, CK-BB, brain - CK-2, CK-MB, heart - CK-3, CK-MM, muscle 38. an elevated CK can indicate what? - MI - Musculare dystrophy - Convulsive seizures - Brain disease---strokes 39. what is the time line (peak and return to normal) of CK in an MI (heart attack)? - Rises 4-6 hrs - Peak at 24 hr - Normal in 3-4 days 40. in the colorimetric method for CK determination, what product is measeure and at what wavelength? - Inorganic phosphorous formed, then quantitated by the Fiske-Subbarow procedure - Inorganic phosphorus is measure at 620 to 700nm ( the amount of phosphorus formed is proportional to CK activity) 41. int eh UV method for CK determination, what product is measure and at what wavelength? - NAADPH at 340 nm 42. in an MI how do the activities of CK, LD, and AST compare? - There is an overall increase in all these enzymes 43. what enzymes are good indicators of pancreatic disease? - Amylase and lipase 44. name the type of amylase produced in humans and state its function - alpha amylase---splits polysaccharide alpha 1-4 linkages in a random manner 45. why doesnt salivary amylase affect the analysis of pancreatic amylase? - Salivary amylase is deactivated by the acid condition of the stomach - Salivary amylase never makes it to the serum - Only serum (pancreatic) amylase is measured 46. elevated serum amylase is indicative of what? - Acute pancreatitis

Carcinoma of the pancrease Mumps Administration of opiates

47. in general, what is the principle of the Cherry-Crandall method of lipase analysis? - Patient serum is incubated with 50% trioloin oil and 5% gum acacia in a pH of 7.0 phosphate buffer - The liberated fatty acid is then quantified by titration against 0.05M NaOH by use of a phenolphthalein indicator 48. what is the clinical significance of elevated lipase activity? - Acute pancreatitis - Pancreatic carcinoma 49. what is the clinical significance of elevated ACP activity? - Carcinoma of the prostate, padgets disease, hyperparathyroidism with skeletal involvement, malignant invasion of the bone, benign hypertrophy of the prostate - Non prostatic ACP---rape cases, Gauchers & Neimann-pick disease, myelocytic leukemia, hematological disorders 50. what is the main diff between ALP & ACP? - The pH - ACP is activated at 5.0 6.0 - ALP is activated at pH of 10.35 51. increased cholinesterase activity is indicative of what? - Can be used to test for liver function - May indicate insecticide poisoning